Marine Biology Research Division, University of California-San Diego, La Jolla, CA 92093-0202, USA.
Environ Microbiol. 2012 Feb;14(2):453-66. doi: 10.1111/j.1462-2920.2011.02633.x. Epub 2011 Nov 8.
Plasmid diversity is still poorly understood in pelagic marine environments. Metagenomic approaches have the potential to reveal the genetic diversity of microbes actually present in an environment and the contribution of mobile genetic elements such as plasmids. By searching metagenomic datasets from flow cytometry-sorted coastal California seawater samples dominated by cyanobacteria (SynMeta) and from the Global Ocean Survey (GOS) putative marine plasmid sequences were identified as well as their possible hosts in the same samples. Based on conserved plasmid replication protein sequences predicted from the SynMeta metagenomes, PCR primers were designed for amplification of one plasmid family and used to confirm that metagenomic contigs of this family were derived from plasmids. These results suggest that the majority of plasmids in SynMeta metagenomes were small and cryptic, encoding mostly their own replication proteins. In contrast, probable plasmid sequences identified in the GOS dataset showed more complexity, consistent with a much more diverse microbial population, and included genes involved in plasmid transfer, mobilization, stability and partitioning. Phylogenetic trees were constructed based on common replication protein functional domains and, even within one replication domain family, substantial diversity was found within and between different samples. However, some replication protein domain families appear to be rare in the marine environment.
在海洋浮游环境中,质粒多样性仍未得到充分了解。宏基因组学方法有可能揭示环境中实际存在的微生物的遗传多样性,以及移动遗传元件(如质粒)的贡献。通过搜索流式细胞术分选的沿海加利福尼亚海水样本的宏基因组数据集(SynMeta)和全球海洋调查(GOS)中可能的海洋质粒序列,以及在同一样本中它们的可能宿主。基于从 SynMeta 宏基因组预测的保守质粒复制蛋白序列,设计了用于扩增一个质粒家族的 PCR 引物,并用于证实该家族的宏基因组 contigs 来自质粒。这些结果表明,SynMeta 宏基因组中的大多数质粒较小且隐匿,主要编码自身的复制蛋白。相比之下,在 GOS 数据集识别的可能质粒序列显示出更大的复杂性,与更具多样性的微生物种群一致,包括涉及质粒转移、移动性、稳定性和分区的基因。基于常见的复制蛋白功能结构域构建了系统发育树,即使在一个复制蛋白家族内,在不同样本内和之间也发现了大量的多样性。然而,一些复制蛋白结构域家族在海洋环境中似乎很少见。